Modules of Antibody Response Profiling
- High throughput antibody response profiling, using thousands of custom peptides
- Verification of the peptide hits for hundreds of samples
- Validation of results by robust custom peptide ELISA
Our Antibody Response Profiling Service Includes
- Evaluation and selection of the optimal strategy for your project
- Design of optimal peptide sequences
- Manufacturing of tailored PepStar™ peptide microarrays and/or custom peptide ELISA plates
- Optimization of arrays and peptides for your requirements
- Incubation & control experiments using your samples
- Detailed data evaluation and analysis supported by our experienced bioinformaticians
- A comprehensive report outlining experiments and results of the study
Module 1: Multiplexed Epitope Discovery
Assay Format: High content peptide microarray
No. of Peptides: Up to 6912 peptides in triplicates per sample
No. of Samples: 1 sample per slide
Principle: Large number of peptides tested against a limited number of samples
Advantages: Low cost per peptide, high peptide throughput
Application: First screening round for the selection of relevant peptides from thousands of candidate peptides
Module 2: Selective Antigen Profiling
Assay Format: Multiwell peptide microarray
No. of Peptides: Up to 188 peptides in triplicates per sample
No. of Samples: 21 samples per slide
Principle: Selected peptides tested against a large number of samples
Advantages: Low cost per sample, high sample throughput
Application: Second screening round to verify candidate peptides for a significant number of samples
Module 3: Marker Validation
Assay Format: Peptide ELISA
No. of Peptides: Up to 96 peptides per plate
No. of Samples: Up to 96 samples per plate
Principle: Flexible for low numbers of peptides & samples
Advantages: Economical common assay
Application: Third screening round for validation of hits or diagnostic assays
Case Study: Epitope Mapping of Epstein-Barr Virus Antigens
High density peptide microarrays have been proven to be efficient tools to map the immune response at a single epitope resolution for large panels of antigens requiring only minute amount of serum or other patient samples (1 µl per assay) to perform a complete analysis [Barouch et al., 2013].
Herein we present a comprehensive but modular humoral immune response profiling platform allowing a systematic and high resolution analysis of the individual antibody repertoire comprising the following techniques:
(1) Multiplexed Epitope Discovery: a limited number of samples (typically 20 to 200) are profiled on PepStar™ High Content Peptide Microarrays displaying up to 6912 peptides in triplicates. This allows the coverage of a broad range of known and potentially relevant cancer antigens as well as antigens from infectious agents.
(2) Selective Antigen Profiling: pre-selected antigens and peptides will be tested against a large number of samples (typically 200 to 1000). For this purpose PepStar™ Multiwell Peptide Microarrays will be used taking advantage of parallel and economic testing of 21 patient samples on a single microarray. The format is limited to 188 candidate peptides (in triplicate) per sample allowing systematic profiling of candidate peptides with large numbers of test samples.
(3) Marker Validation: a robust Peptide ELISA platform was developed based on highly purified peptides allowing both, the validation of the selected candidates as well as detailed analysis of the recognition. The platform enables transfer of results towards diagnostic assays and the selection of vaccine candidates.
The performance of this new platform is exemplarily demonstrated for selecting relevant epitopes from a large panel of antigens of the Epstein-Barr Virus.
Read more in our Application Note.
Quality Assurance
All production is ISO 9001:2015 certified.
